Confirmation of Germ-Line Transmission in the Red Fluorescence Protein (RFP) Transgenic Cloned Male Cat

2010 ◽  
Vol 12 (6) ◽  
pp. 739-747 ◽  
Author(s):  
Su-Jin Cho ◽  
Young S. Lee ◽  
Jae-Ik Lee ◽  
Jae-Il Bang ◽  
Jing Yang ◽  
...  
Keyword(s):  
Germ Line ◽  
Red Fluorescence ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein ◽  
Germ Line Transmission

302 CONFIRMATION OF GERM-LINE TRANSMISSION OF RED FLUORESCENCE PROTEIN TRANSGENIC MALE CAT

2009 ◽  
Vol 21 (1) ◽  
pp. 248 ◽  
Author(s):  
S. J. Cho ◽  
Y. S. Lee ◽  
E. G. Choi ◽  
J. I. Bang ◽  
X. F. Yu ◽  
...  
Keyword(s):  
Germ Line ◽  
Basic Research ◽  
Transgenic Animals ◽  
Pcr Analysis ◽  
Parental Line ◽  
Tissue Samples ◽  
Red Fluorescence ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein ◽  
Natural Mating

Production of transgenic animals is highly desirable for biotechnology and basic research. Therefore, a method of producing genetically modified cats through genetic engineering is important for generating biomedical models of human diseases. We investigate reproductive ability of red fluorescence protein (RFP) transgenic cloned male cat (RFP TG cat) in natural mating with domestic female cat. One domestic female cat in natural mating with RFP TG cat delivered 6 kittens of which 3 (2 female and 1 male) showed RFP expression of parental line. Among the 3 RFP expressing kittens, 1 died at day 5 and were subjected to PCR analysis for confirmation of RFP gene expression. Red fluorescence protein gene was detected in tissue samples including liver, muscle, brain, large intestine, bladder, spleen, sexual organ, small intestine, kidney, heart, stomach, pancreas, lungs, and skin of the kitten. These results indicated that RFP transgenic cloned male cat have normal reproductive fertility, and the stability of transmission of the RFP transgene through the germ line and its normal expression in offspring. Supported by KOSEF (M10525010001-05N2501-00110). S. J. Cho, Y. S. Lee, E. G. Choi, and J. I. Bang were supported by a scholarship from the Post BK21 Program, the Ministry of Education, Science and Technology, Korea.

scholarly journals The Exosporium of Bacillus megaterium QM B1551 Is Permeable to the Red Fluorescence Protein of the Coral Discosoma sp.

2016 ◽  
Vol 7 ◽  
Author(s):  
Mariamichela Lanzilli ◽  
Giuliana Donadio ◽  
Roberta Addevico ◽  
Anella Saggese ◽  
Giuseppina Cangiano ◽  
...  
Keyword(s):  
Bacillus Megaterium ◽  
Red Fluorescence ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein

56 ESTABLISHMENT OF TRANSGENIC RED FLUORESCENCE PROTEIN (RFP) CLONE DOGS THROUGH A STABLE TRANSMISSION OF RFP GENE TO NEXT GENERATION

2011 ◽  
Vol 23 (1) ◽  
pp. 134
Author(s):  
H. J. Oh ◽  
J. E. Park ◽  
M. J. Kim ◽  
G. A. Kim ◽  
E. J. Park ◽  
...  
Keyword(s):  
Second Generation ◽  
Pcr Analysis ◽  
Next Generation ◽  
Wild Type ◽  
Red Fluorescence ◽  
Reproductive Ability ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein ◽  
Regulated Gene Expression ◽  
Stable Transmission

Somatic cell nuclear transfer (SCNT) technology has been spotlighted not only for its advantage in producing unlimited numbers of genetically identical animals, but also the possibility of producing complex genetic modifications in animals. However, a few reports showed that mosaic expression of transgene in transgenic animals produced by SCNT (Park et al. 2002) and down-regulated gene expression is sometimes irreversible in their offspring (Bordignon et al. 2003). Therefore, we investigated reproductive ability by a breeding between female transgenic beagles and wild-type beagles. When female transgenic beagles (R1, R2, R3, and R5) expressing red fluorescence protein (RFP) gene reached puberty at 373, 353, 283, and 354 days after birth, serum progesterone concentration was monitored for detecting timing of ovulation. Approximately 72 to 79 h after ovulation, the beagles were naturally mated or artificially inseminated. Pregnancy was confirmed by ultrasonography at Day 30 after insemination. The transgenic bitches (R1, R2, R3, and R5) were then bred with wild-type male dogs, became pregnant, and successfully delivered 13 puppies (9 female and 4 male). In order to prove integration of RFP gene in all offspring, DNA was extracted from the blood of pups on Day 7 after birth. For PCR analysis, a primer pair for the RFP gene, forward primer (5′CGTGAAGCTGAAGGTGA-3′) and reverse primer (5′-CTCGTACTGCTCCACGA-3′), were used to amplify a 517-bp DNA fragment. The initial denaturation was performed at 94°C for 5 min, followed by 30 cycles at 94°C for 40 s (denaturation), 58°C for 40 s (annealing), and 72°C for 40 s (extension), and a final incubation at 72°C for 10 min to ensure complete strand extension. Presence of the RFP transgene in 7 of the puppies was confirmed by PCR and the puppies expressed RFP upon UV illumination. It was not different from the 53.8% expected Mendelian ratio. The present result demonstrated a stable transmission of the RFP gene into 5 female and 2 male offspring in the second generation. Among the second generation, 2 female puppies integrated with the RFP gene were in heat at ∼1-year-old. They were then bred with the semen of a wild-type beagle and bore 6 puppies. In the third generation, 3 puppies carried the RFP gene and results showed the expected Mendelian ratio. In conclusion, the present study demonstrates that female transgenic beagles have normal reproductive ability and a stable insertion of the transgene to the next generation. This study was financially supported by NRF (#M10625030005-508-10N25), SNU foundation (Benefactor; RNL BIO), BK 21 for Veterinary Science, and Purina Korea.

Coral red fluorescence protein as genetic modified baculovirus tracer

2005 ◽  
Vol 119 (3) ◽  
pp. 255-259 ◽  
Author(s):  
Tzyy-Rong Jinn ◽  
Suey-Sheng Kao ◽  
Jason T.C. Tzen ◽  
Tzong-Yuan Wu
Keyword(s):  
Red Fluorescence ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein

32 NEURON-SPECIFIC EXPRESSION OF THE RED FLUORESCENCE PROTEIN IN CLONED DOGS

2012 ◽  
Vol 24 (1) ◽  
pp. 128 ◽  
Author(s):  
H. J. Oh ◽  
J. E. Park ◽  
M. J. Kim ◽  
G. Kim ◽  
E. J. Park ◽  
...  
Keyword(s):  
Southern Blot ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Degenerative Disease ◽  
Neural Cells ◽  
Red Fluorescence ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein ◽  
The Brain ◽  
Synapsin 1

The pathogenesis of neuronal degenerative disease as Alzheimer's disease (AD) has been a subject of intensive research for the last few decades worldwide. But despite such effort, treatment or preventive measures for AD have so far made no breakthrough. One of the contributing factors that hindered the progress of research is the lack of appropriate AD models. Mouse models have limitations for AD research because the irreconcilable species gap between the rodent and human has impeded the research itself as well as the application of the findings from the rodent studies to human cases. As an alternative, here we performed a preliminary study to develop novel neuronal degenerative disease models using a canine transgenic somatic cell nuclear transfer (SCNT) technology. The aim of this study is to produce a transgenic dog that expresses neuron-specific transgene in the brain by SCNT. In this study, we chose human synapsin 1 promoter as primarily neuron selective, driving the red fluorescent protein transgene. For SCNT, synapsin 1-red fluorescence protein (SYN1-RFP) was introduced into female beagle adipose-derived stem cell via lentiviral vector infection. The SYN1-RFP cells were injected into enucleated in vivo-matured dog oocytes and fused by electric stimulation. The fused couplets (80/94, 85.1%) were chemically activated and transferred into the uterine tube of 5 naturally oestrus-synchronized surrogates. Three of them (60%) maintained pregnancy and subsequently gave birth to 3 cloned pups (SYN1-RFP A, SYN1-RFP B, SYN1-RFP C) by natural delivery or cesarean section. Birth weights of the offspring ranged from 120 to 280 g and SYN1-RFP C is still alive, healthy and does not show any abnormalities. The microsatellite analysis shows that all SYN1-RFP puppies originated from the SYN1-RFP cells used in SCNT and mitochondrial DNA analysis shows that the puppies had been derived from the oocyte donors. In order to investigate the result in multiple transgene insertions, SYN-RFP puppies were screened by Southern blot analysis using DNA extracted from skin biopsies. Transgene copy number was estimated by Southern blot analysis. The SYN-RFP A and B that died at 3 days after birth had approximately 5 and 2 copies of the transgene integrated, respectively, whereas the alive SYN-RFP C has 1 copy. SYN-RFP B was particular in that it did not express RFP in the entire body, but samples collected postmortem showed expression of the RFP transgene under the human synapsin 1 promoter in neural cells in the brain of SYN-RFP B. In conclusion, we report here that (1) the human synapsin promoter is functional in neural cells of dog brain and (2) a neural-specific-transgene-expressed dog was generated for the first time by transgenic SCNT technique. Furthermore, the SYN-RFP dog has great potential to understand the function of a neuronal degenerative disease model dog. This study was supported by MKE (Grant # 10033839-2011-13), RNL Bio, IPET and TS Corporation.

scholarly journals Non-invasive molecular and functional imaging of cytosine deaminase and uracil phosphoribosyltransferase fused with red fluorescence protein

2008 ◽  
Vol 47 (7) ◽  
pp. 1211-1220 ◽  
Author(s):  
Ligang Xing ◽  
Xuelong Deng ◽  
Khushali Kotedia ◽  
Ellen Ackerstaff ◽  
Vladimir Ponomarev ◽  
...  
Keyword(s):  
Functional Imaging ◽  
Cytosine Deaminase ◽  
Red Fluorescence ◽  
Non Invasive ◽  
Uracil Phosphoribosyltransferase ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein

Generation of a recloned transgenic cat expressing red fluorescence protein

2010 ◽  
Vol 73 (7) ◽  
pp. 848-855 ◽  
Author(s):  
S.J. Cho ◽  
J.I. Bang ◽  
X.F. Yu ◽  
Y.S. Lee ◽  
J.H. Kim ◽  
...  
Keyword(s):  
Red Fluorescence ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein
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